SG OneStep PRO RT-qPCR kit is one-step RT-qPCR kit that provides accurate real-time quantification of RNA targets. The kit consists of unique, proofreading thermostable reverse transcriptase and DNA Polymerase in an easy-to-use format.
The kit provides:
- 2x PRO RT-qPCR SG Buffer that is a universal reaction buffer with dNTPs (dTTP is partially replaced with dUTP) that can be used on most real-time PCR cyclers available. Allows as option to use thermolabile uracil-N-glycosylase (UNG).
- The SG PRO Enzyme Mix contains unique highly sensitive and thermostable reverse transcriptase and DNA Polymerase, RNase Inhibitor and SYBR Green I dye.
- Reverse transcriptase works in a wide temperature range from 52-72°C without losing specificity and sensitivity.
- Both cDNA synthesis and PCR are performed in a single tube using gene-specific primers and either total RNA or mRNA.
- Proofreading is maintained during both reverse transcription and PCR steps.
- SYBR Green I is a fluorescent dye which binds all doublestranded DNA molecules and emits a fluorescent signal on binding. The excitation and emission maxima of SYBR Green I are at 494 nm and 521 nm, respectively, which are compatible with any real-time cycler.
- Kit contains thermolabile uracil-N-glycosylase (UNG) that is optimized for RT-qPCR reactions.
- If cyclers from Applied Biosystems are used ROX passive reference dye is necessary. SG OneStep RT-qPCR kit is provided in two variants: without ROX and with ROX Solution provided separately. The table below shows recommended amount of ROX (25 µM) required for a specific PCR cycler.